Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8

Joon Ho Jang, Jin Seok Kim, Hyung Chan Seo, Seung Lee Tae, Yoo Hong Min, Jee Sook Hahn, Yun Woong Ko

Research output: Contribution to journalArticle

Abstract

IL-8 secreted from human umbilical vein endothelial cells(HUVEC) is known to induce apoptosis of myelogeneous leukemia cell line. Clostridium difficile(C. difficile) toxin A promotes IL-8 secretion from a large variety of cell types, including monocy tes and intestinal epithelial cells but such effect on endothelial cells is unknown. In this study, we investigated whether C. difficile toxin A could induce IL-8 secretion from HUVEC, and the resultant effect on apoptosis of leukemia cell lines. For the detection of IL-8 in culture media, an enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used. After 3Id passage, HUVEC were cocultured with leukemia cell lines(K562, U937, KG1; 1 x 10s cells/mL) with or without C.difficile toxin A(lmg/ml) in 24-well plates. After 2 days, the supernatants were collected by centrifugation at 15,000 rpm and were applied to the ELIS A kit. Leukemia cells and supernatants were collected for the detection of apoptosis and 1L8 level, respectively. Cell counting, Wright-Giemsa staining, and Annexin V-PI apoptosis kit were used for the detection of apoptosis and enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used for the detection of IL-8 in supernatants. As a control, leukemia cell lines were cultured with HUVEC. All experiments were performed triplicate. The levels of IL-8 in supernatants of each cell lines were higher when C.difficile toxin A was added; 65.4 ±4.8 ng/ml to 80.7 ±5.2 ng/ml(in K562), 59.6 ±3.3 ng/ml to 76.2 ±4.6 ng/ml(in U937), 70.6 ±2.4 ng/ml to 85.7 ±6.1 ng/ml(in KG 1 ). The levels of apoptosis were also increased in samples cocultured with C.difficile toxin A; 21.4 ±1.7% to 40.2 ± 2.1 %(in K562), 20.6 ±2.4% to 49.6 ±2.9%(in U937), 22.7 ±1.9% to 47.6 ±3.1 %(in KG1 ). When leukemia cell lines cultured only with C.difficile toxin A in the absence of HUVEC, the levels of IL-8 in supernatants and the levels of apoptosis of each cell lines showed similar results to the samples cocultured without C.difficile toxin A. This study showed that C. difficile toxin A could induce apoptosis against leukemic cells, possibly by promoting secretion of IL-8 from vascular endothelium.

Original languageEnglish
Pages (from-to)202b-203b
JournalBlood
Volume96
Issue number11 PART II
Publication statusPublished - 2000 Dec 1

Fingerprint

Clostridium
Clostridium difficile
Interleukin-8
Up-Regulation
Endothelial Cells
Apoptosis
Endothelial cells
Human Umbilical Vein Endothelial Cells
Cells
Leukemia
Cell Line
Immunosorbents
Assays
Enzyme-Linked Immunosorbent Assay
Centrifugation
Annexin A5
Vascular Endothelium
Enzymes
Culture Media
Epithelial Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Jang, J. H., Kim, J. S., Seo, H. C., Lee Tae, S., Min, Y. H., Hahn, J. S., & Ko, Y. W. (2000). Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8. Blood, 96(11 PART II), 202b-203b.
Jang, Joon Ho ; Kim, Jin Seok ; Seo, Hyung Chan ; Lee Tae, Seung ; Min, Yoo Hong ; Hahn, Jee Sook ; Ko, Yun Woong. / Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8. In: Blood. 2000 ; Vol. 96, No. 11 PART II. pp. 202b-203b.
@article{b856acdfe3144b42a3a5ab6b4030e437,
title = "Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8",
abstract = "IL-8 secreted from human umbilical vein endothelial cells(HUVEC) is known to induce apoptosis of myelogeneous leukemia cell line. Clostridium difficile(C. difficile) toxin A promotes IL-8 secretion from a large variety of cell types, including monocy tes and intestinal epithelial cells but such effect on endothelial cells is unknown. In this study, we investigated whether C. difficile toxin A could induce IL-8 secretion from HUVEC, and the resultant effect on apoptosis of leukemia cell lines. For the detection of IL-8 in culture media, an enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used. After 3Id passage, HUVEC were cocultured with leukemia cell lines(K562, U937, KG1; 1 x 10s cells/mL) with or without C.difficile toxin A(lmg/ml) in 24-well plates. After 2 days, the supernatants were collected by centrifugation at 15,000 rpm and were applied to the ELIS A kit. Leukemia cells and supernatants were collected for the detection of apoptosis and 1L8 level, respectively. Cell counting, Wright-Giemsa staining, and Annexin V-PI apoptosis kit were used for the detection of apoptosis and enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used for the detection of IL-8 in supernatants. As a control, leukemia cell lines were cultured with HUVEC. All experiments were performed triplicate. The levels of IL-8 in supernatants of each cell lines were higher when C.difficile toxin A was added; 65.4 ±4.8 ng/ml to 80.7 ±5.2 ng/ml(in K562), 59.6 ±3.3 ng/ml to 76.2 ±4.6 ng/ml(in U937), 70.6 ±2.4 ng/ml to 85.7 ±6.1 ng/ml(in KG 1 ). The levels of apoptosis were also increased in samples cocultured with C.difficile toxin A; 21.4 ±1.7{\%} to 40.2 ± 2.1 {\%}(in K562), 20.6 ±2.4{\%} to 49.6 ±2.9{\%}(in U937), 22.7 ±1.9{\%} to 47.6 ±3.1 {\%}(in KG1 ). When leukemia cell lines cultured only with C.difficile toxin A in the absence of HUVEC, the levels of IL-8 in supernatants and the levels of apoptosis of each cell lines showed similar results to the samples cocultured without C.difficile toxin A. This study showed that C. difficile toxin A could induce apoptosis against leukemic cells, possibly by promoting secretion of IL-8 from vascular endothelium.",
author = "Jang, {Joon Ho} and Kim, {Jin Seok} and Seo, {Hyung Chan} and {Lee Tae}, Seung and Min, {Yoo Hong} and Hahn, {Jee Sook} and Ko, {Yun Woong}",
year = "2000",
month = "12",
day = "1",
language = "English",
volume = "96",
pages = "202b--203b",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "11 PART II",

}

Jang, JH, Kim, JS, Seo, HC, Lee Tae, S, Min, YH, Hahn, JS & Ko, YW 2000, 'Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8', Blood, vol. 96, no. 11 PART II, pp. 202b-203b.

Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8. / Jang, Joon Ho; Kim, Jin Seok; Seo, Hyung Chan; Lee Tae, Seung; Min, Yoo Hong; Hahn, Jee Sook; Ko, Yun Woong.

In: Blood, Vol. 96, No. 11 PART II, 01.12.2000, p. 202b-203b.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8

AU - Jang, Joon Ho

AU - Kim, Jin Seok

AU - Seo, Hyung Chan

AU - Lee Tae, Seung

AU - Min, Yoo Hong

AU - Hahn, Jee Sook

AU - Ko, Yun Woong

PY - 2000/12/1

Y1 - 2000/12/1

N2 - IL-8 secreted from human umbilical vein endothelial cells(HUVEC) is known to induce apoptosis of myelogeneous leukemia cell line. Clostridium difficile(C. difficile) toxin A promotes IL-8 secretion from a large variety of cell types, including monocy tes and intestinal epithelial cells but such effect on endothelial cells is unknown. In this study, we investigated whether C. difficile toxin A could induce IL-8 secretion from HUVEC, and the resultant effect on apoptosis of leukemia cell lines. For the detection of IL-8 in culture media, an enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used. After 3Id passage, HUVEC were cocultured with leukemia cell lines(K562, U937, KG1; 1 x 10s cells/mL) with or without C.difficile toxin A(lmg/ml) in 24-well plates. After 2 days, the supernatants were collected by centrifugation at 15,000 rpm and were applied to the ELIS A kit. Leukemia cells and supernatants were collected for the detection of apoptosis and 1L8 level, respectively. Cell counting, Wright-Giemsa staining, and Annexin V-PI apoptosis kit were used for the detection of apoptosis and enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used for the detection of IL-8 in supernatants. As a control, leukemia cell lines were cultured with HUVEC. All experiments were performed triplicate. The levels of IL-8 in supernatants of each cell lines were higher when C.difficile toxin A was added; 65.4 ±4.8 ng/ml to 80.7 ±5.2 ng/ml(in K562), 59.6 ±3.3 ng/ml to 76.2 ±4.6 ng/ml(in U937), 70.6 ±2.4 ng/ml to 85.7 ±6.1 ng/ml(in KG 1 ). The levels of apoptosis were also increased in samples cocultured with C.difficile toxin A; 21.4 ±1.7% to 40.2 ± 2.1 %(in K562), 20.6 ±2.4% to 49.6 ±2.9%(in U937), 22.7 ±1.9% to 47.6 ±3.1 %(in KG1 ). When leukemia cell lines cultured only with C.difficile toxin A in the absence of HUVEC, the levels of IL-8 in supernatants and the levels of apoptosis of each cell lines showed similar results to the samples cocultured without C.difficile toxin A. This study showed that C. difficile toxin A could induce apoptosis against leukemic cells, possibly by promoting secretion of IL-8 from vascular endothelium.

AB - IL-8 secreted from human umbilical vein endothelial cells(HUVEC) is known to induce apoptosis of myelogeneous leukemia cell line. Clostridium difficile(C. difficile) toxin A promotes IL-8 secretion from a large variety of cell types, including monocy tes and intestinal epithelial cells but such effect on endothelial cells is unknown. In this study, we investigated whether C. difficile toxin A could induce IL-8 secretion from HUVEC, and the resultant effect on apoptosis of leukemia cell lines. For the detection of IL-8 in culture media, an enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used. After 3Id passage, HUVEC were cocultured with leukemia cell lines(K562, U937, KG1; 1 x 10s cells/mL) with or without C.difficile toxin A(lmg/ml) in 24-well plates. After 2 days, the supernatants were collected by centrifugation at 15,000 rpm and were applied to the ELIS A kit. Leukemia cells and supernatants were collected for the detection of apoptosis and 1L8 level, respectively. Cell counting, Wright-Giemsa staining, and Annexin V-PI apoptosis kit were used for the detection of apoptosis and enzyme-linked immunosorbent assay(ELISA) system for human IL-8 was used for the detection of IL-8 in supernatants. As a control, leukemia cell lines were cultured with HUVEC. All experiments were performed triplicate. The levels of IL-8 in supernatants of each cell lines were higher when C.difficile toxin A was added; 65.4 ±4.8 ng/ml to 80.7 ±5.2 ng/ml(in K562), 59.6 ±3.3 ng/ml to 76.2 ±4.6 ng/ml(in U937), 70.6 ±2.4 ng/ml to 85.7 ±6.1 ng/ml(in KG 1 ). The levels of apoptosis were also increased in samples cocultured with C.difficile toxin A; 21.4 ±1.7% to 40.2 ± 2.1 %(in K562), 20.6 ±2.4% to 49.6 ±2.9%(in U937), 22.7 ±1.9% to 47.6 ±3.1 %(in KG1 ). When leukemia cell lines cultured only with C.difficile toxin A in the absence of HUVEC, the levels of IL-8 in supernatants and the levels of apoptosis of each cell lines showed similar results to the samples cocultured without C.difficile toxin A. This study showed that C. difficile toxin A could induce apoptosis against leukemic cells, possibly by promoting secretion of IL-8 from vascular endothelium.

UR - http://www.scopus.com/inward/record.url?scp=33748528101&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748528101&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:33748528101

VL - 96

SP - 202b-203b

JO - Blood

JF - Blood

SN - 0006-4971

IS - 11 PART II

ER -

Jang JH, Kim JS, Seo HC, Lee Tae S, Min YH, Hahn JS et al. Clostridium difficile toxin a induces apoptosis in leukemic cells by upregulation of endothelial il-8. Blood. 2000 Dec 1;96(11 PART II):202b-203b.